LSM 5 PASCAL Manual

8-S. LSM 5 PASCAL Englisch
01.10.1999
15:42 Uhr
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Microscopy from Carl Zeiss
LSM 5 PASCAL
The Personal
Laser Scanning Microscope
Focused on Your Success.
8-S. LSM 5 PASCAL Englisch
01.10.1999
15:42 Uhr
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LSM 5 PASCAL
An LSM of the Fifth Generation
Your work involves the acquisition of two-dimen-
Then you should consider a very personal laser
sional and three-dimensional images? And you ex-
scanning microscope:
pect optimum quality along with high resolution?
The LSM 5 PASCAL – a laser scanning microscope
You also need to record time series? You want to
of the fifth generation which has been designed
acquire the image data in different orientations
for you or your team working in routine and
and reconstruct at different viewing angles with-
research.
out the need to remount your specimen? And you
especially expect to be able to perform quanti-
The LSM 5 PASCAL is the ‘little brother’ of the
tative measurements of, for example, calcium
leading edge LSM 510 system.
concentrations and pH-values, areas and surface
‘Little’, however, is only descriptive of its ama-
properties?
zingly attractive price, as our engineers have made
no compromises to image quality, sensitivity and
flexibility!
More than 150 years of innovation in optics and
around 20 years of experience in all fields of laser
scanning microscopy combined with the ongoing
dialog with you, the users, make the LSM 5
PASCAL a rewarding long-term investment. An
investment which does not put a strain on your
budget and which can be expanded as your
Detector
applications requirements change or grow.
Pinhole
in Confocal
Plane
Beam Splitter
Laser
Scan Mirror
xy
Objective
Specimen
Focal Plane
z
The LSM 5 PASCAL represents
a compact system that uses
confocal technique and the
point scanning method to acquire
3 dimensional pictures of your
specimen with the highest resolution,
close to physical limitations.
8-S. LSM 5 PASCAL Englisch
01.10.1999
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LSM 5 PASCAL
Yours Personally
Maximum operating convenience...
...highest resolution...
Like all LSM units of the fifth generation, the hall-
In addition, the LSM 5 PASCAL provides image
mark of LSM 5 PASCAL is a fully motorized
sizes of up to 2,048x2,048 pixels, scan fields
scanning module. All moving components, such
of unrivaled size along with maximum linearity,
as emission filter wheels, main and secondary
12-bit resolution per channel, high scan speeds,
dichroic beam splitters, the pinhole and the
high-sensitivity detectors (selected photomulti-
mechanical attenuators for each laser line, are
pliers) and short light paths from the specimen to
computer-controlled and take up the required po-
the detector – features which are unique in its
sition making the set-up error free and without
class.
any need for cumbersome manual operation. A
special benefit in practice: the position can be
...and great flexibility.
reset automatically at the press of a button.
Special emphasis has been placed on the flexibility of the LSM 5 PASCAL. Six versions with one
or two confocal channels and combinations of
lasers in the blue and green spectrum and a choice
of five microscope stands are available for most
varied applications in the life and materials sciences. The LSM 5 PASCAL can be converted from an
“The LSM 5 PASCAL grows with
the needs of your applications:
You can add a second fluorescence channel as well as a transmitted light detector. Up to
8 single emission filters per
channel and the six position
dichroic mirror wheels are
exchangeable.
Furthermore you can chose from
a variety of software options.”
upright to an inverted microscope within minutes.
Confocal microscopy is no more than a flick of the
wrist away from conventional microscopy.
3/1
Xanthidium cristatum (SAG 173.80),
Immunofluorescence chloroplasts (red),
storage vesicle (green) + DIC overlay
(Lab for Experimental Phycology,
Göttingen)
3
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8-S. LSM 5 PASCAL Englisch
01.10.1999
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8-S. LSM 5 PASCAL Englisch
01.10.1999
15:43 Uhr
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LSM 5 PASCAL
Proven Software
on a New Platform
A system is always only as good as its user inter-
In addition to data acquisition, the software of the
face. For this reason, the LSM 5 PASCAL uses
LSM 5 PASCAL provides many 2D and 3D presen-
proven technology as its basis: the fast and reliable
tation options, numerous on-line and off-line
software runs on a high-end PC under Windows
measuring functions and a very convenient image
NT and is closely related to the tried and tested
database for the management and the documen-
software of the LSM 510. Just like the configura-
tation of the images.
tion of the microscope and the scan module, the
software functionality can also be extended step
The software controls the motorized system
by step.
components, i.e. the microscope, the scan and
the laser modules. The user-friendly and intuitive
The synchronized control of scanners, data acqui-
user interface makes your daily work easier. As all
sition and input/output signals by a digital signal
adjustable components are controlled by the scan
processor (DSP) allows extremely flexible scan
and laser module software, system configurations
strategies.
once set can be reactivated by simply pressing a
button, time-consuming manual settings are now
Various scan functions are available for data
a thing of the past. Macros can be used for re-
acquisition. They include multifluorescence ima-
peating individual work processes.
ges free from cross talk and the scanning of any
number of regions of interest (ROI) of almost all
forms. The acquisition of 3D stacks combined with
time series supports the researcher studying processes in living cells.
6/1:
“The Crop function:
Fast and easy selection
of a new scan area”
OK cells, mitosis, eCFP indicates
PSD95 and Alexa546 shows Actin
(Dr. Klöcker, Uni Konstanz)
6/2:
“Defining and modifying
multiple arbitrary Regions
Of Interest (ROI)”
Cells, MnSOD shown using Cy2,
GFAP indicated by Cy3, 2kx2k pixels
(Dr. Possel, Institute of Medical
Neurobiology, Magdeburg)
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Benefits for your applications:
• Reproducible experiments by a simple mouse click
• Large scan fields and high resolution images up to
2,048x2,048 pixels
• Flexible scan strategies also for 3D and time series
• Continuously adjustable pinhole for optimum z resolution
• Precisely controlable laser intensities to protect your
specimens and to reduce the bleaching of dyes
• Multifluorescence images without cross talk between
the channels
7/7
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LSM 5 PASCAL
Specification
7/3
Microscopes
z Drives
Fine focusing stage
XY stage
Scan Module
7/5
Scan resolution
Scan speed
Scan zoom
Scan rotation
Scan field
Pinhole
Detection
Dynamic range
Laser module
Laser lines
Attenuation
Electronic’s module
7/6
LSM 5 Control
Computer
Monitor
Software
Upright: Axioplan 2 MOT, Axioskop 2 MOT, Axioskop 2 FS MOT,
Inverted: Axiovert 100 M Side/BasePort
Accessories like ICS objectives and filter cubes can be used
DC servo or step motor, smallest steps starting at 25 nm
HRZ 200, total range 200 µm, smallest step 10 nm
Motorized XY scanning stage, smallest step 250 nm
Two independent galvo scanning mirrors
1x4 to 2048x2048 pixels, user definable
2x10 levels, line frequencies from 4 up to 1300 Hz; 0.4 s per frame 512x512 pixels
0.7x to 8x, variable with steps of 0.1
Any angle, variable with steps of 1°
18 mm diagonal in the primary image plane (with zoom factor 1x)
One pinhole with variable diameter size, adjustable
1 or 2 confocal R/FL channels with built in highly sensitive photomultiplier tubes
1 external transmitted light channel (DIC capable) optional
12 bit per channel
Ar laser 488, 514 nm, 25 mW - HeNe laser 543 nm, 1 mW
(end of lifetime specifications)
Individual and variable intensity control of all lines
Control circuitry for microscope, laser and scan module
with built in high performance Digital Signal Processor (DSP)
Well equipped High-end PC with ample RAM + hard disk space, many accessories,
multi user operating system Windows NT 4.0
Ergonomic high contrast large screen monitor, 2 nd monitor optional
Software for operating microscope, laser module and scan module;
For image acquisition, display, processing and archiving, line and frame scan, 3D or/and time
lapse recording, Multitracking and dual direction scanning, spline scanning, ROIs
(region of interest), 3D projections, quantitative measurements, many options, more
than 20 export file formats (LSM, TIF, BMP, JPG, PCX, GIF, …)
Some of the components and software functions are optional.
7/1
C. elegans with embryos,
autofluorescence + DIC,
(single frame out of time series),
(Sample courtesy of Dr. Afaq, MBL
Woods Hole)
7/2
Fly eye (Musca domesticus), double
fluorescence, 3D reconstruction,
(Dr. Lam, Dr. Jans, J. Curtin School
of Medical Research, Sydney)
7/3
Zebrafish embryo, neurons (green),
NCAM (red)
(Dr. Marx, Dr. Bastmeyer, Uni Konstanz)
7/6
Fetal small intestine, double fluorescence,
stereo projection
(Dr. Hashimoto, Jikei University, Tokyo)
7/4
HT29 cells loaded with
Fluo-4, Line scan time series, 60 min
(Dr. Nitschke, Uni Freiburg)
7/7
Drosophila brain, neural circuit indicated with GFP
(Dr. Ito, NI of Basic Biology Lab, Okasaki)
7/5
Triticum spelta, auto fluorescence,
maximum projection
7/8
Endothelial cells, Texas Red marks F-actin (red),
BODIPY shows tubulin (green)
multitracking scan without crosstalk of channels
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Carl Zeiss
Microscopy
D-07740 Jena
Phone: ++49-36 41/64 -1616
Telefax: ++49-36 41/64 -3144
E-mail: micro@zeiss.de
Internet: www.zeiss.de/micro
Subject to change.
7/8
Printed on environment-friendly paper,
bleached without the use of chlorine.
For further details, please contact:
40-053 e/ 09.99